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[用于鉴定从血培养中分离出的酵母的商业多重串联聚合酶链反应方法的评估]

[Evaluation of a Commercial Multiplex Tandem Polymerase Chain Reaction Method for the Identification of the Yeasts Isolated from Blood Cultures].

作者信息

Er Halil, Koyuncu Özyurt Özlem, Özhak Betil, Yazısız Hatice, Öngüt Gözde, Eres Sarıtaş Zübeyde, Dönmez Levent, Çolak Dilek, Günseren Filiz, Öğünç Dilara

机构信息

Health Sciences University, Antalya Training and Research Hospital, Clinic of Medical Microbiology, Antalya, Turkey.

Akdeniz University Faculty of Medicine, Department of Medical Microbiology, Antalya, Turkey.

出版信息

Mikrobiyol Bul. 2020 Oct;54(4):596-605. doi: 10.5578/mb.69832.

DOI:10.5578/mb.69832
PMID:33107288
Abstract

Candidemia is one of the most important health care-associated infections worldwide. Candida species have species-specific antifungal susceptibility profiles and it has been shown that the identification of the Candida species is necessary for the appropriate treatment of the patients with candidemia. Various methods are used to shorten the identification time for the determination of the causative species. Fungal ID multiplex tandem polymerase chain reaction (MT-PCR) (AusDiagnostics, Australia) is a test developed to identify yeasts and molds isolated from clinical specimens. In this study, we aimed to evaluate the Fungal ID MT-PCR test (AusDiagnostics, Australia) for the identification of the yeasts from positive blood cultures in Akdeniz University Hospital Central Laboratory. Between December 2016 and December 2017, blood culture samples from 92 consecutive patients with yeast cells detected in Gram stained smears were tested by Fungal ID MT-PCR and the reference method. After the subculture of the positive signaling blood culture bottles to Sabouraud dextroz agar (SDA), the identification of the yeasts were performed by morphological identification methods (Germ tube test, Corn Meal Tween® 80 agar media, etc.), BD Phoenix Yeast ID Panel (Becton Dickinson, Sparks, MD) and Bruker Biotyper matrix-assisted laser desorption ionization-time of mass spectrometry (MALDI-TOF MS) (Bruker Daltonics, Germany) systems. Identification with MALDI-TOF MS have been accepted as the reference method. Thirty-five of the isolates were identified as Candida albicans, 17 were Candida glabrata, 13 were Candida parapsilosis, 12 were Candida tropicalis, seven were Candida krusei , two were Candida guilliermondii, two were Candida dubliniensis, two were Candida inconspicua, one was Candida kefyr and one was Saprochaete capitata by the reference method. In our study, no blood culture sample yielded more than one yeast species. 94.6% of the strains were presumptively identified by the morphological identification methods. Discordant results were not detected between the BD Phoenix Yeast ID Panel and the reference method. Thirty-three of the isolates were identified as C.albicans, 15 were C.glabrata, 13 were C.parapsilosis, 11 were C.tropicalis, five were C.krusei , two were C.guilliermondii, one was C.dubliniensis, one was C.kefyr and 10 were Candida spp. by Fungal ID MT-PCR assay. Since C.inconspicua and S.capitata were not included in the test panel, C.inconspicua was identified as Candida spp. in two samples, while S.capitata could not be identified in one sample. Concordance between Fungal ID MT-PCR and the reference method were found to be 88% at the species level and 98.9% at the genus level. The sensitivity of the Fungal ID MT-PCR test in in the detection of C.krusei and C.glabrata was 71.4% and 88.2%, respectively. Fungal ID MT-PCR test has shown a high performance in the identification at the genus level, but the identification at the species level, which is important for the treatment management, was moderate. Fungal ID MT-PCR can be used as an adjunct test to the traditional identification methods for the early identification of the Candida species.

摘要

念珠菌血症是全球最重要的医疗保健相关感染之一。念珠菌属具有种特异性的抗真菌药敏谱,并且已经表明,对于念珠菌血症患者的恰当治疗,鉴定念珠菌属是必要的。为缩短确定致病菌种的鉴定时间,人们采用了各种方法。真菌鉴定多重串联聚合酶链反应(MT-PCR)(澳大利亚AusDiagnostics公司)是一种用于鉴定从临床标本中分离出的酵母和霉菌的检测方法。在本研究中,我们旨在评估真菌鉴定MT-PCR检测法(澳大利亚AusDiagnostics公司)在阿克德尼兹大学医院中心实验室对阳性血培养物中的酵母进行鉴定的效果。在2016年12月至2017年12月期间,对92例在革兰氏染色涂片上检测到酵母细胞的连续患者的血培养样本,采用真菌鉴定MT-PCR和参考方法进行检测。将发出阳性信号的血培养瓶接种到沙氏葡萄糖琼脂(SDA)上进行传代培养后,通过形态学鉴定方法(芽管试验、玉米粉吐温80琼脂培养基等)、BD Phoenix酵母鉴定板(美国BD公司,斯帕克斯,马里兰州)和布鲁克微生物鉴定系统基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)(德国布鲁克道尔顿公司)系统对酵母进行鉴定。采用MALDI-TOF MS进行鉴定被视为参考方法。通过参考方法鉴定出35株白色念珠菌、17株光滑念珠菌、13株近平滑念珠菌、12株热带念珠菌、7株克柔念珠菌、2株季也蒙念珠菌、2株都柏林念珠菌、2株 inconspicua念珠菌、1株凯菲念珠菌和1株头状腐质霉。在我们的研究中,没有血培养样本分离出一种以上的酵母菌种。94.6%的菌株通过形态学鉴定方法得到初步鉴定。BD Phoenix酵母鉴定板与参考方法之间未检测到不一致的结果。通过真菌鉴定MT-PCR检测法鉴定出33株白色念珠菌、15株光滑念珠菌、13株近平滑念珠菌、11株热带念珠菌、5株克柔念珠菌、2株季也蒙念珠菌、1株都柏林念珠菌、1株凯菲念珠菌和10株念珠菌属。由于检测板中未包括 inconspicua念珠菌和头状腐质霉,在两个样本中 inconspicua念珠菌被鉴定为念珠菌属,而在一个样本中头状腐质霉无法鉴定。真菌鉴定MT-PCR与参考方法在菌种水平的一致性为88%,在属水平的一致性为98.9%。真菌鉴定MT-PCR检测法对克柔念珠菌和光滑念珠菌的检测灵敏度分别为71.4%和88.2%。真菌鉴定MT-PCR检测法在属水平的鉴定中表现出较高的性能,但在对治疗管理很重要的菌种水平的鉴定中表现中等。真菌鉴定MT-PCR可作为传统鉴定方法的辅助检测手段,用于念珠菌属菌种的早期鉴定。

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